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心房肌高效的基因转染是一项技术难题.心房血供少、心房壁薄等原因限制了心房肌基因转染方法的发展.但心肌基因治疗作为一种研究和治疗心脏疾病的有效方法而逐渐被学者重视.本研究探讨一种经心房外膜涂抹使携带有绿色荧光蛋白(EGFP)基因的腺病毒高效靶向转染心房肌细胞方法的可行性.经犬胸骨正中开胸术,将含有一定浓度携带EGFP基因的腺病毒、胰蛋白酶和poloxamer F407的混合溶液涂抹至右心房外膜.术后经荧光显微镜观察荧光强度和实时PCR检测EGFP mRNA表达水平.EGFP荧光强度和mRNA表达水平在第1～6周呈现先增高后减低的趋势,其高峰在第3周出现;第3周右心房前壁各层心肌细胞均有EGFP表达;右心耳EGFP mRNA表达水平高于右心房前壁,右心房前壁和后壁表达水平无明显差异;房间隔的表达水平低于右心房,但明显高于左心房;心室和其它脏器如肺脏、肝脏、脾脏、骨骼肌、胃壁及小肠壁等的表达水平远低于右心房;通过HE染色及Masson’s染色,第3周心肌无明显炎症反应和纤维化改变.因此,经心房外膜涂抹的方法使基因靶向转染心肌细胞具有较好的高效性、靶向性和安全性.
Atrial muscle efficient gene transfection is a technical problem.A small amount of atrial blood supply, atrial thin wall and other factors restricting the development of atrial gene transfection method.But myocardial gene therapy as a study and treatment of heart disease and effective method Which is paid more and more attention by scholars.This study is to explore the feasibility of transfection of atrial adenocarcinoma carrying green fluorescent protein (EGFP) A mixture of adenovirus, trypsin and poloxamer F407 containing a certain concentration of EGFP gene was applied to the right atrial appendage.Fluorescence intensity and real-time PCR were used to detect the expression of EGFP mRNA.Fluorescence intensity and mRNA expression level of EGFP From the first week to the sixth week, the first increase and then the decrease appeared. The peak appeared at the third week. In the third week, the expression of EGFP in each layer of right atrial anterior wall was higher than that in the right atrium. There was no significant difference in the expression level of the anterior and posterior walls of the right atrium; the atrial septum was lower than the right atrium but significantly higher than the left atrium; the ventricles and other organs such as lung, liver and spleen The expression level of skeletal muscle, stomach wall and small intestine wall was much lower than that of the right atrium. There was no obvious inflammatory reaction and fibrosis in myocardium by HE staining and Masson’s staining at the third week.Therefore, Transfection of myocardial cells with good efficiency, targeting and safety.